Atomic resolution crystal structures, EXAFS, and quantum chemical studies of rusticyanin and its two mutants provide insight into its unusual properties

Rusticyanin from the extremophile Thiobacillus ferrooxidans is a blue copper protein with unusually high redox potential and acid stability. We present the crystal structures of native rusticyanin and of its Cu site mutant His143Met at 1.27 and 1.10 A, respectively. The very high resolution of these structures allows a direct comparison with EXAFS data and with quantum chemical models of the oxidized and reduced forms of the proteins, based upon both isolated and embedded clusters and density functional theory (DFT) methods. We further predict the structure of the Cu(II) form of the His143Met mutant which has been experimentally inaccessible due to its very high redox potential. We also present metrical EXAFS data and quantum chemical calculations for the oxidized and reduced states of the Met148Gln mutant, this protein having the lowest redox potential of all currently characterized mutants of rusticyanin. These data offer new insights into the structural factors which affect the redox potential in this important class of proteins. Calculations successfully predict the structure and the order of redox potentials for the three proteins. The calculated redox potential of H143M ( approximately 400 mV greater than native rusticyanin) is consistent with the failure of readily available chemical oxidants to restore a Cu(II) species of this mutant. The structural and energetic effects of mutating the equatorial cysteine to serine, yet to be studied experimentally, are predicted to be considerable by our calculations.     

Bioenergy crop models: descriptions,data requirements,and future challenges

Field studies that address the production of lignocellulosic biomass as a source of renewable energy provide critical data for the development of bioenergy crop models. A literature survey revealed that 14 models have been used for simulating bioenergy crops including herbaceous and woody bioenergy crops, and for crassulacean acid metabolism (CAM) crops. These models simulate field‐scale production of biomass for switchgrass (ALMANAC, EPIC, and Agro‐BGC), miscanthus (MISCANFOR, MISCANMOD, and WIMOVAC), sugarcane (APSIM, AUSCANE, and CANEGRO), and poplar and willow (SECRETS and 3PG). Two models are adaptations of dynamic global vegetation models and simulate biomass yields of miscanthus and sugarcane at regional scales (Agro‐IBIS and LPJmL). Although it lacks the complexity of other bioenergy crop models, the environmental productivity index (EPI) is the only model used to estimate biomass production of CAM (Agave and Opuntia) plants. Except for the EPI model, all models include representations of leaf area dynamics, phenology, radiation interception and utilization, biomass production, and partitioning of biomass to roots and shoots. A few models simulate soil water, nutrient, and carbon cycle dynamics, making them especially useful for assessing the environmental consequences (e.g., erosion and nutrient losses) associated with the large‐scale deployment of bioenergy crops. The rapid increase in use of models for energy crop simulation is encouraging; however, detailed information on the influence of climate, soils, and crop management practices on biomass production is scarce. Thus considerable work remains regarding the parameterization and validation of process‐based models for bioenergy crops; generation and distribution of high‐quality field data for model development and validation; and implementation of an integrated framework for efficient, high‐resolution simulations of biomass production for use in planning sustainable bioenergy systems.     

Distribution and genotypic characterization of Salmonella serovars isolated from tropical seafood of Cochin, India

Aims:  To determine the distribution of Salmonella serovars in seafood and to examine the intraserovar genetic variations in Salmonella enterica subsp. enterica serovar Rissen and Salmonella Weltevreden by polymerase chain reaction (PCR)-ribotyping and enterobacterial repetitive intergenic consensus (ERIC)-PCR methods.
Method and Results:  A total of 417 seafood samples collected over 2003–2006 from fishing harbours and fish markets of Cochin (India) was studied for presence of Salmonella serovars. Seafood samples were analysed for the presence of Salmonella by Bacteriological Analytical Manual (BAM), U.S. Food & Drug Administration (USFDA) method. The study indicated that 23·2% of the seafood samples were positive for Salmonella and a total of 241 Salmonella isolates comprising of 27 different serovars were isolated from seafood. S. Weltevreden, Salmonella Rissen, Salmonella Typhimurium and Salmonella Derby were found to be the most predominant serovars in seafood. PCR-ribotypes and ERIC-PCR profiles showed multiple genotypic profiles for S. Rissen and S. Weltevreden in seafood and the level of discrimination indices obtained was at 0·974 for S. Rissen and 0·988 for S. Weltevreden, respectively.
Conclusion:  The study highlighted the major Salmonella serovars in the seafood of Cochin (India) and molecular fingerprinting pattern revealed genetic variation among S. Rissen and S. Weltevreden.
Significance and Impact of the Study:  Widespread occurrence of Salmonella contamination in seafood and multiple clones of S. Rissen and S. Weltevreden detected in seafood, thus, indicated the diverse routes of Salmonella contamination in seafood.     

Anopheles culicifacies Y-chromosome dimorphism indicates sibling species (B and E) with different malaria vector potential in Sri Lanka

In Sri Lanka, malaria is transmitted mainly by Anopheles culicifacies Giles sensu lato (Diptera: Culicidae). In India, this nominal taxon comprises sibling species A, B, C, D and E, distinguished by their chromosome morphology. Species B (identified by polytene chromosome sequence Xab, 2g1 + h1) is not such an efficient vector of malaria as other members of the An. culicifacies complex in India. All specimens of An. culicifacies s.l. examined from Sri Lanka possess Xab, 2g1 + h1 polytenes, previously interpreted as species B, despite their important vector status. Recently, species E was described from Rameshwaram Island (Tamil Nadu, India) between Sri Lanka and the Indian mainland, where both species B and E are sympatric. Species B and E share polytene sequence Xab, 2g1 + h1 but differ by the mitotic Y-chromosome being acrocentric in species B, submetacentric in species E, the latter implicated as vector of vivax malaria. From May 1999 to January 2000, we surveyed Y-chromosomes of male progeny from An. culicifacies Xab, 2g1 + h1 females collected from cattle bait in diverse malarious districts of Sri Lanka: Badulla, Monaragala, Puttalam and Trincomalee. Karyotypes of readable quality were obtained from 42/83 families examined, with overall proportions 24% acrocentric and 76% submetacentric Y-chromosome carriers, both types being sympatric in at least 3/4 localities sampled. By analogy with the situation on Rameshwaram Island, we interpret these observations to demonstrate widespread presence of two members of the An. culicifacies complex in Sri Lanka, their karyotypes being compatible with species B and E, the latter predominant and having greater vector potential.     

Altered expression of neuronal nitric oxide synthase in the duodenum longitudinal muscle-myenteric plexus of obesity induced diabetes mouse: implications on enteric neurodegeneration

Type 2 diabetes caused by obesity shows autonomic neuropathy. Molecular mechanism involved in enteric neurodegeneration is not clear. Neuronal nitric oxide synthase (nNOS) is one of the important agents involved in gastrointestinal function. Therefore, expression of nNOS in the duodenum LM-MP of type 2 diabetes model mouse was studied. Real time RT-PCR analysis showed reduction in nNOS expression in male diabetic LM-MP compared to male control. In contrast, female diabetic LM-MP had high level of nNOS mRNA compared to female control. Western blot determination of LM-MP showed reduction in nNOS protein in male diabetic LM-MP and high level of nNOS in female diabetic LM-MP compared to their respective controls. Expression of nNOS observed by Western blot was further confirmed by nNOS immunostaining of the mouse duodenum. TUNEL staining of mouse duodenum showed apoptosis in male diabetic enteric neurons. These studies suggest that nNOS expression in LM-MP varies with gender during early stage of type 2 diabetes. In addition, reduced expression of nNOS is likely to contribute to apoptosis seen in the enteric neurons of male type 2 diabetic mice.     

Isolation,expansion and characterisation of mesenchymal stem cells from human bone marrow,adipose tissue,umbilical cord blood and matrix: a comparative study

The multipotent and immunosuppressive capacities of mesenchymal stem cells (MSCs) attract several scientists worldwide towards translational research focusing on treatment of diseases including liver failure. Though MSC’s have been isolated from different sources, researchers do not concur on the best source for expansion and clinical translation. In this study, we have compared the isolation, proliferation and expansion of MSCs from umbilical cord blood (UCB), Wharton’s Jelly (WJ), bone marrow (BM) and adipose tissue (AT). MSCs were isolated by density gradient separation from UCB, BM and AT and by both enzymatic and explant method for WJ. The MSCs are characterized by their ability to adhere to plastic, expression of positive (CD105, CD73, CD90, CD29, CD44) and negative (CD45, CD14, CD34) markers by flow cytometry and also by their in vitro adipogenic, osteogenic and chondrogenic differentiation. This comprehensive study clearly shows that WJ is better than UCB both in terms of rapidity, yield and ease of procedure. AT and BM are autologous sources for MSC’s but the specimen collection involves cumbersome and painful procedures and an invasive approach. However being autologous, they are safe and probable candidates for therapeutic future applications.

Electronic supplementary material

The online version of this article (doi:10.1007/s10616-014-9718-z) contains supplementary material, which is available to authorized users.     

Electrospun Carbon Nanofibers Encapsulated with NiCoP: A Multifunctional Electrode for Supercapattery and Oxygen Reduction,Oxygen Evolution,and Hydrogen Evolution Reactions

Functionalizing nanostructured carbon nanofibers (CNFs) with bimetallic phosphides enables the material to become an active electrode for multifunctional applications. A facile electrospinning technique is utilized for the first time to develop NiCoP nanoparticles encapsulated CNFs that are used as an energy storage system of supercapattery, and as an electrocatalyst for oxygen reduction, oxygen evolution, and hydrogen evolution reaction in KOH electrolyte. Evolving from the inclusion of bimetallic phosphide nanoparticles, the NiCoP/CNF electrode unveils superior‐specific capacitance (333 Fg^?1 at 2 Ag^?1) and rate capability (87%). The fabricated supercapattery device offers a voltage of 1.6 V that supplies a remarkable energy density (36 Wh kg^?1) along with an improved power density (4000 W kg^?1) and unwavering cyclic stability (25 000 cycles). Meanwhile, the NiCoP/CNF electrode has simultaneously performed well as a multifunctional electrocatalyst for oxygen reduction reaction at a half‐wave potential of 0.82 V versus reversible hydrogen electrode and can attain a current density of 10 mA cm^?2 at a very low overpotential of 268 and 130 mV for the oxygen evolution reaction and hydrogen evolution reaction, respectively. Thus, the NiCoP/CNF with all its inimitable electrode properties has profoundly proved its proficiency at handling multifunctional challenges in terms of both storage and conversion.